PROJECT SUMMARY Targeted delivery of therapeutic gene editing CRISPR ribonucleoprotein (RNP) complexes to cells offers major opportunities for next-generation therapies, however, realization of this goal requires a vehicle to protect and direct CRISPR RNPs to their targeted cells and tissues. Currently, there are no vehicles that can incorporate CRISP RNPs for cellular delivery in vivo. Here, we propose to use structured DNA nanoparticles (DNA-NPs) as versatile carriers for the targeted delivery of RNPs, taking advantage of base-pairing between the RNP and the DNA-NP to facilitate RNP incorporation and controlled release. DNA-NPs will be designed with stoichiometric control over the number of RNP payloads they carry internally, and functionalized externally with antibodies and lipid binding moieties to control cellular targeting and intracellular trafficking. We will minimize macrophage engulfment and enhance blood compatibility through surface functionalization with polyethylene glycol (PEG) and other biocompatible passivation techniques, using standard assays for nanoparticle physiological and immunological interactions. Binding and release of CRISPR RNPs from DNA-NPs will be assayed in vitro, and DNA-RNP base pairing will ensure stable binding and facile release of cargo from vehicle. Genome editing will be assayed through both next generation sequencing and targeted PCR, which will esure that the enzymatic activity of the cargo remains intact during delivery. Targeting CRISPR RNPs to sub-populations of cultured cells expressing cell specific surface receptors will be evaluated, with particular attention paid targeted gene editing in cells of interest. The generality of our RNP delivery platform offers a unique and transformative approach to treating a range of deadly and currently untreatable genetic diseases and cancers by targeted delivery of CRISPR.